DAPI, permeability - (Jun/10/2008 )

Hi, can DAPI enter live cells?According to invitrogen, DAPI only stains dead cells. However, wikipedia says DAPI stains both live/dead cells.Also, are there any other molecules (fluorescent or nonfluorescent) that binds to cell nucleus and are membrane impermeant?

-liumaikuh-

QUOTE (liumaikuh @ Jun 10 2008, 10:15 PM)

Hi, can DAPI enter live cells?According to invitrogen, DAPI only stains dead cells. However, wikipedia says DAPI stains both live/dead cells.Also, are there any other molecules (fluorescent or nonfluorescent) that binds to cell nucleus and are membrane impermeant?

live cell staining of nucleus is an interesting quesrtion; I tried Hoe 33342 which should be used for live cell staining of nucleus, but I did not get any staining;

I never tried DAPI for living cells; I suppose it will arrest cell cycle;

if you try DAPI for live cells please let us now about your results

-The Bearer-

QUOTE (The Bearer @ Jun 11 2008, 02:46 PM)

QUOTE (liumaikuh @ Jun 10 2008, 10:15 PM)

Hi, can DAPI enter live cells?According to invitrogen, DAPI only stains dead cells. However, wikipedia says DAPI stains both live/dead cells.Also, are there any other molecules (fluorescent or nonfluorescent) that binds to cell nucleus and are membrane impermeant?

live cell staining of nucleus is an interesting quesrtion; I tried Hoe 33342 which should be used for live cell staining of nucleus, but I did not get any staining;

I never tried DAPI for living cells; I suppose it will arrest cell cycle;

if you try DAPI for live cells please let us now about your results

I don`t remember exactly if it was DAPI or Hoechst but a colleague tried to do ON- live cell imaging on stained cells but she did not get nice movies as cells were detaching and also dying after a while.

you may or may not find what you need here - Molecular Probes has a number of kits for distinguishing live vs. dead cells and you can probably answer your questions about dyes that can permeate the nucleus of a live cell, if you look around this link

-aimikins-

You can express GFP-Histone (not sure which, but it is published) and make some fantastic movies of cell division.

-rosewater-

QUOTE (rosewater @ Jun 13 2008, 01:28 PM)

You can express GFP-Histone (not sure which, but it is published) and make some fantastic movies of cell division.

nice idea! do you know anything about cell cycle or proliferation as GFP-histone may have epigenetic effects

-The Bearer-

QUOTE (The Bearer @ Jun 13 2008, 01:50 PM)

QUOTE (rosewater @ Jun 13 2008, 01:28 PM)

You can express GFP-Histone (not sure which, but it is published) and make some fantastic movies of cell division.

nice idea! do you know anything about cell cycle or proliferation as GFP-histone may have epigenetic effects

I know a bit. But the histone may be an odd one, like H2AX, which might matter. My Research Associate made such a transfectant in my last lab to look at mitotic defects in response to drugs. In an inducible vector, I believe. The main problem with GFP in general (for time lapse photography) is that exposing the cells to UV for long periods is not good.

-rosewater-

I use DAPI to stain live cells which are hard to destinguis seperately (e.g. fused myocytes), works great.

-Freebird-

QUOTE (Freebird @ Jun 17 2008, 05:52 AM)

I use DAPI to stain live cells which are hard to destinguis seperately (e.g. fused myocytes), works great.

great! how to perform? DAPI in PBS? which concentration of DAPI is good? how long do you incubate with DAPI?